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小鼠Aire逆转录病毒表达载体的构建及表达
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作者 李娟 张君 张毓 《天津医科大学学报》 2011年第4期443-446,463,共5页
目的:将小鼠Aire基因定向连入MigR1-GFP质粒,得到长期稳定的Aire逆转录病毒表达载体,收集逆转录病毒上清感染胸腺基质细胞,为研究Aire在胸腺细胞终末分化中的作用奠定基础。方法:以小鼠胸腺基质细胞的cDNA为模板扩增得到Aire的ORF片段;... 目的:将小鼠Aire基因定向连入MigR1-GFP质粒,得到长期稳定的Aire逆转录病毒表达载体,收集逆转录病毒上清感染胸腺基质细胞,为研究Aire在胸腺细胞终末分化中的作用奠定基础。方法:以小鼠胸腺基质细胞的cDNA为模板扩增得到Aire的ORF片段;将该片段连接入MigR1-GFP质粒;将构建成功的Aire-MigR1-GFP质粒转染293T细胞,收集逆转录病毒上清;使用逆转录病毒上清感染胸腺基质细胞系MTEC9,流式细胞仪检测感染效率,Western blot确定Aire蛋白的表达。结果:(1)成功得到Aire的ORF片段。(2)构建的Aire逆转录病毒表达质粒能够成功表达Aire蛋白。(3)逆转录病毒上清成功感染胸腺基质细胞系MTEC9。结论:成功构建了小鼠Aire逆转录病毒表达载体,并实现了Aire蛋白在胸腺基质细胞系中的强制性表达。 展开更多
关键词 小鼠 AIRE MigR1 胸腺基质细胞
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EXAMPLES OF CLINICAL APPLICATION OF ACUPUNCTURE OF ZHONGWAN(CV12) WITH ELONGATED NEEDLE
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作者 吴建华 阎莉 《World Journal of Acupuncture-Moxibustion》 2005年第1期43-45,共3页
In the present paper, the authors in troduce Professor YAN Li’s experie nce on clinical application of elongated needle for the treatment of gastralgia, migraine and hysteria by acupuncture of Zhongwan (中脘 CV 12). ... In the present paper, the authors in troduce Professor YAN Li’s experie nce on clinical application of elongated needle for the treatment of gastralgia, migraine and hysteria by acupuncture of Zhongwan (中脘 CV 12). In addition, the y also introduce its operation methods and some points for attention. During ope ration of the elongated needle, the operator should manipulate it gently, avoidi ng rude insertion and rash withdrawal; rather, utilize the elasticity of the steel wire to press the needle downward slowly, and cause the favorable needlin g sensations to spread slowly downward to the affected region along with the mov ement of the needle body. 展开更多
关键词 Elongated needle Clinical application Gastralgia Migr aine Hysteria
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利用MigR1质粒构建CD19-CAR逆转录病毒载体优化其对人原代T细胞的转染效率 被引量:3
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作者 王扬 唐古生 +9 位作者 徐丽丽 阮婕 程辉 周虹 华亦斐 胡晓霞 顾海慧 钱宝华 王健民 杨建民 《中华血液学杂志》 CAS CSCD 北大核心 2015年第4期331-336,共6页
目的优化二代CD19嵌合抗原受体(CD19-CAR)逆转录病毒载体对人原代T细胞的转染效率。方法应用重组DNA技术将CD19.CAR构建人逆转录病毒载体,转入plat-A包装细胞收集病毒上清并转染人原代T细胞及K562细胞系,采用流式细胞术检测不同条... 目的优化二代CD19嵌合抗原受体(CD19-CAR)逆转录病毒载体对人原代T细胞的转染效率。方法应用重组DNA技术将CD19.CAR构建人逆转录病毒载体,转入plat-A包装细胞收集病毒上清并转染人原代T细胞及K562细胞系,采用流式细胞术检测不同条件下的转染效率,用RT-PCR方法检测目的基因的表达情况,用ELISA法检测IFN-γ、TNF-α表达水平。结果①MigR1-CD19-CAR逆转录病毒载体成功构建后包装高滴度逆转率病毒;②相同条件下,MigR1-CD19-CAR逆转录病毒对K562细胞系转染效率显著高于T细胞;③对T细胞,离心120min组的转染率为(54.5±14.6)%,明显高于未离心组、离心30min组及离心60min组(P值均〈0.05);④CD3/CD28磁珠与rhIL-2联合对人T细胞活化扩增效率个体差异显著,根据扩增倍数优化个体转染时机可提高转染效率(最高达69.3%),且RT-PCR检测证实CD19.CAR目的基因在转染后72hT细胞中高效特异性转录;⑤CD19-CAR-T细胞与CD19.K562细胞共培养组IFN-γ、TNF-α表达水平均增加,分别为(13230±1543)pg/ml、(4217±211)pg/ml,与阴性对照组比较差异均有统计学意义(P值均〈0.01)。结论通过MigR1-CD19-CAR逆转录病毒载体转染人原代T细胞,并通过延长离心转染时间、根据扩增倍数确定个体转染时机而成功优化转染效率,RT-PCR检测CD19-CAR目的基因在转染后人T细胞中高效特异性转录,CD19-CAR-T细胞特异性识别靶细胞引发IFN-γ、TNF-α表达水平增加。 展开更多
关键词 受体 抗原 质粒 MigR1 转染 T淋巴细胞 细胞因子类
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Russian Physicians in Harbin (1920-32)
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作者 Pavel Ratmanov Yan Liu Fengmin Zhang 《Frontiers of History in China》 2019年第3期353-384,共32页
This study analyzes medical practitioners’adaptation to a dynamic cultural and political scene and examines the impact of medical refugees on a local community.In the early 1920s,there was an influential Russian medi... This study analyzes medical practitioners’adaptation to a dynamic cultural and political scene and examines the impact of medical refugees on a local community.In the early 1920s,there was an influential Russian medical community in Harbin that established medical societies and medical schools.The organization of medical societies was a part of the active formation of a professional community and represented a thoughtful|measure for countering the control of Chinese officials.The high degree of cooperation between Russian and Chinese medical personnel in the medical-sanitary department of the Chinese Eastern Railway and in Harbin municipal medical facilities was a part of Harbin physicians’activities. 展开更多
关键词 RUSSIA modern China Chinese EASTERN RAILWAY Manchuria HARBIN medical refugees émigré PHYSICIANS Soviet influence
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Reservoir formation conditions and key technologies for exploration and development in Shengtuo oilfield in Bohai Bay Basin
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作者 Mingshui Song 《Petroleum Research》 2020年第4期289-303,共15页
The Shengtuo oilfield is the largest monolithic oilfield in the Bohai Bay Basin and even in the continental fault basins of Eastern China.Since discovered in 1963,the Shengtuo oilfield experiences the early exploratio... The Shengtuo oilfield is the largest monolithic oilfield in the Bohai Bay Basin and even in the continental fault basins of Eastern China.Since discovered in 1963,the Shengtuo oilfield experiences the early exploration stage of rapid reserve discovery and production,the middle exploration stage of high and stable production and development,and the late exploration stage of theoretical and technological innovation for expanding the field of exploration,research on fine development technology of high-water-cut reservoir for enhanced oil recovery,and maintaining slow decline of oil production.Re-serves of this oilfield increase in each stage.By the end of 2018,after more than 50 years of exploration and development,the Shengtuo oilfeld has cumulative proven oil reserves of 0.512 x 10°t,cumulative oil production of 0.193 x 109t,and it keeps stable annual production of more than 150 x 105t in 2018.Abundant hydrocarbon resources,large anticline structure near to oil source,multiple types of reservoirs and traps,and favorable hydrocarbon migration channels provide favorable hydrocarbon accumulation conditions for the Shengtuo oilfield.Fine development techniques such water-flooding in the early development stage,fine reservoir description in the high water cut stage,stratigraphic subdivision and well pattem vector adjustment,tapping remaining oil in rhythmic layers,etc,as well as conti nuous progress in exploration technologies of glutenite and turbidite reservoirs around the main body of the oilfield in the high-level exploration stage,provide effective supports for continuous increase of reserves and long-term high and stable production of the Shengtuo oilfeld. 展开更多
关键词 Continental rift basin Hydrocarbon acumulation conditions Source-reservoir-caprodk combinations Hydrocarbon migr ation dhannels Key technologies for exploration and DEVELOPMENT Shengtuo oilfield Bohai Bay Basin
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